Project Summary The goal of this SBIR project is to design and develop an efficient, automation- compatible Directed Cell Migration (DCM) 96-well Assay that distinguishes chemokinetic from chemotactic behavior. Cell migration is currently assayed using trans-membrane well inserts, microfluidic systems and various versions of scratch assays. These methods are expensive and provide inconsistent data. Due to its importance in cancer and associated metastatic processes, cell migration is intensively studied and in vitro cell migration assays are routinely used in drug discovery programs aimed at identifying new cancer therapeutics. An important characteristic of cancer cells is their ability to detect chemoattractant gradients and migrate in response to them. A major barrier to efficient screening of candidate cancer therapeutics affecting directed cell migration is the lack of affordable cell based assays that are robust, physiologically relevant, reproducible and cost-effective to perform. Platypus Technologies LLC has developed and commercialized two distinct products for the study of cell migration. The Oris DCM assay proposed in this Phase I project builds on the success of the Platypus Oris Pro cell migration assay product. Specifically, the Oris DCM assay will allow measurement of directed cell migration in response to chemoattractants, thereby facilitating identification and development of lead therapeutic compounds that target cellular responses to chemoattractants. The proposed assay is amenable to automation and to readout by microscopy, multi-well plate readers, and high content imagers. Major additional benefits of the proposed assay format include ability to 1) visualize cell movement in real-time and 2) perform multiplexed secondary screens to elucidate mechanism of action of the test compound via cytostaining within the same well that the primary chemotaxis assay was run. This latter benefit is an extraordinary advantage that increases the knowledge-generating power of the research dollar by conserving reagents including compounds, cells, etc and resources that would otherwise be consumed in repetitive testing required by competing formats.